Gebski Bijanka L, Errington Stephen J, Johnsen Russell D, Fletcher Susan, Wilton Stephen D
Experimental Molecular Medicine Group, Centre for Neuromuscular and Neurological Disorders, University of Western Australia, Nedlands, Perth 6097, Western Australia.
Neuromuscul Disord. 2005 Oct;15(9-10):622-9. doi: 10.1016/j.nmd.2005.06.009.
Induction of specific exon skipping during the processing of the dystrophin gene transcript is being pursued as a potential therapy for Duchenne muscular dystrophy. Antisense oligonucleotides directed at motifs involved in pre-mRNA processing can manipulate dystrophin exon incorporation in the mature gene transcript. We have compared the exon skipping ability of oligodeoxyribonucleotides with compounds of the identical sequence incorporating 2'-O-methyl modified bases. Antisense oligonucleotides composed entirely of 2'-O-methyl modified bases on a phosphorothioate backbone were consistently more efficient at inducing exon skipping than comparable oligodeoxyribonucleotides. Chimeric antisense oligonucleotides, mixtures of unmodified and 2'-O-methyl modified bases, induced intermediate levels of exon skipping. In addition, we describe terminal modifications that may be incorporated into the 2'-O-methyl antisense oligonucleotides to further enhance efficiency of exon skipping. Our findings suggest that 2'-O-methyl antisense oligonucleotides should be considered for human clinical trials involving targeted exon skipping in dystrophin gene expression in preference to oligodeoxyribonucleotides.
在肌营养不良蛋白基因转录本加工过程中诱导特定外显子跳跃，正作为杜氏肌营养不良症的一种潜在治疗方法而被探索。针对参与前体mRNA加工的基序的反义寡核苷酸，可以操纵成熟基因转录本中肌营养不良蛋白外显子的掺入。我们比较了寡脱氧核糖核苷酸与掺入2'-O-甲基修饰碱基的相同序列化合物的外显子跳跃能力。在硫代磷酸酯主链上完全由2'-O-甲基修饰碱基组成的反义寡核苷酸，在诱导外显子跳跃方面始终比可比的寡脱氧核糖核苷酸更有效。嵌合反义寡核苷酸，即未修饰和2'-O-甲基修饰碱基的混合物，诱导中等水平的外显子跳跃。此外，我们描述了可以掺入2'-O-甲基反义寡核苷酸以进一步提高外显子跳跃效率的末端修饰。我们的研究结果表明，在涉及肌营养不良蛋白基因表达中靶向外显子跳跃的人体临床试验中，应优先考虑2'-O-甲基反义寡核苷酸而非寡脱氧核糖核苷酸。
